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Production of knockout rats using ENU mutagenesis and a yeast-based screening assay.

The rat is a widely used model in biomedical research and is often the preferred rodent model in many areas of physiological and pathobiological research. Although many genetic tools are available for the rat, methods to produce gene-disrupted knockout rats are greatly needed. In this study, we developed protocols for creating N-ethyl-N-nitrosourea (ENU)-induced germline mutations in several rat strains. F1 preweanling pups from mutagenized Sprague Dawley (SD) male rats were then screened for functional mutations in Brca1 and Brca2 using a yeast gap-repair, ADE2-reporter truncation assay. We produced knockout rats for each of these two breast cancer suppressor genes.

Pubmed ID: 12754522

Authors

  • Zan Y
  • Haag JD
  • Chen KS
  • Shepel LA
  • Wigington D
  • Wang YR
  • Hu R
  • Lopez-Guajardo CC
  • Brose HL
  • Porter KI
  • Leonard RA
  • Hitt AA
  • Schommer SL
  • Elegbede AF
  • Gould MN

Journal

Nature biotechnology

Publication Data

June 30, 2003

Associated Grants

  • Agency: NCI NIH HHS, Id: CA28954
  • Agency: NCI NIH HHS, Id: CA77494

Mesh Terms

  • Animals
  • BRCA1 Protein
  • BRCA2 Protein
  • Cells, Cultured
  • Ethylnitrosourea
  • Gene Expression Regulation
  • Gene Targeting
  • Male
  • Mutagenesis, Site-Directed
  • Protein Engineering
  • Rats
  • Rats, Mutant Strains
  • Rats, Sprague-Dawley
  • Recombinant Proteins
  • Yeasts